Wang Q, O'Brien P J, Chen C X, Cho D S, Murray J M, Nishikura K
Wistar Institute and Department of Pharmacology, University of Pennsylvania, Philadelphia 19104, USA.
J Neurochem. 2000 Mar;74(3):1290-300. doi: 10.1046/j.1471-4159.2000.741290.x.
Different isoforms of serotonin subtype 2C receptor (5-HT(2C)R) with altered G protein-coupling efficacy are generated by RNA editing, which converts genomically encoded adenosine residues into inosines. In combination, editing of five sites all located within the second intracellular loop region of 5-HT(2C)R mRNA changes the gene-encoded Ile, Asn, and Ile at positions 156, 158, and 160, respectively. We analyzed the G protein-coupling functions of previously unreported editing isoform receptors. An approximately 13-fold reduction in the agonist potency for G protein-coupling stimulation as well as a significantly reduced basal level activity was observed with the thalamus-specific isoform carrying Ile156, Gly158, and Val160 (5-HT(2C)R-IGV). In contrast, the agonist was four- to five-fold less potent with 5-HT(2C)R-MSV and -IDV, detected in the amygdala and choroid plexus, respectively, indicating a dominant role for the amino acid residue at position 158 in receptor functions. We also identified a splicing variant receptor with a truncated C terminus that displayed no ligand binding capacity or G protein-coupling activity. Examination of the alternatively spliced RNA encoding this truncated receptor suggests that editing of this variant RNA occurs after completion of splicing, resulting in complete editing at all five sites.
5-羟色胺2C受体(5-HT(2C)R)的不同亚型通过RNA编辑产生,其G蛋白偶联效率发生改变,RNA编辑可将基因组编码的腺苷残基转变为肌苷。5-HT(2C)R mRNA的五个编辑位点均位于第二个细胞内环区域,编辑后分别改变了基因编码的第156、158和160位的异亮氨酸、天冬酰胺和异亮氨酸。我们分析了之前未报道的编辑亚型受体的G蛋白偶联功能。携带异亮氨酸156、甘氨酸158和缬氨酸160的丘脑特异性亚型(5-HT(2C)R-IGV)对G蛋白偶联刺激的激动剂效力降低了约13倍,基础水平活性也显著降低。相比之下,分别在杏仁核和脉络丛中检测到的5-HT(2C)R-MSV和5-HT(2C)R-IDV的激动剂效力低4至5倍,表明第158位氨基酸残基在受体功能中起主导作用。我们还鉴定出一种C末端截短的剪接变体受体,其不显示配体结合能力或G蛋白偶联活性。对编码这种截短受体的可变剪接RNA的检测表明,这种变体RNA的编辑在剪接完成后发生,导致所有五个位点都发生完全编辑。